The principal objective of the proposed research is to contribute to an understanding of the relationship between the regulation of renal metabolism and renal function. The effects of various hormones and 3'-5' cyclic AMP on the rates of gluconeogenesis and transport processes in isolated tissue slices and tubules, will be examined. These alterations will be compared with changes in the levels of activity and degree of phosphorylation of gluconeogenic enzymes and with the activity of protein kinases and phosphoprotein phosphatases. An important aspect of this proposal will be the correlation of renal metabolism and transport with the phosphorylation of endogenous proteins under various conditions followed by the isolation and identification of these proteins from both the cytosol and membrane fractions of kidney homogenates. The approach will also involve studies with isolated enzymes and reconstructed multienzyme systems. The role of enzyme interactions and binding to subcellular membranes on the regulation of enzymes and proteins involved in gluconeogenesis and transport processes will be examined. The physiological effects of the subcellular distribution of protein kinases on the regulation and coordination of these processes in renal tissue will be evaluated. These studies will utilize a physiological approach to evaluate the effects observed "in vitro" with well-defined isolated enzyme systems to determine whether these effects can also be demonstrated with perfused organs maintained with various hormones under glycolytic or gluconeogenic conditions. It is important to determine whether the regulatory effects seen with isolated enzymes have any role in the control of these enzymes "in vivo".